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Reference Icon Redbook Fluorescent Protein Guide: Empty Backbones


Background

Fluorescent_Cells.jpg

Fluorescent proteins are genetically encoded tools that are used extensively by life scientists. The original green fluorescent protein (GFP) was cloned in 1992 (Prasher et al., Gene, 1992), and since then scientists have engineered numerous GFP-variants and non-GFP proteins that result in a diverse set of colors.

Addgene has assembled a collection of empty plasmid backbones with different fluorescent tags for you to create fusion proteins with your gene of interest. These are organized By Color and By Species Expressed In.

Unsure of which fluorescent protein to use in your experiment? Check out our blog article entitled "Which Fluorescent Protein Should I Use?".

Empty Backbones for Fluorescent Protein Fusions (Organized by Color)


Blue/UV

Protein Excitation Emission Brightness* Structure Plasmids
Sirius 355 424 4 Monomer
SBFP2 380 446 13 Monomer
Azurite 383 450 14 Monomer
EBFP2 383 448 17 Monomer
mKalama1 385 456 16 Monomer
mTagBFP2 399 456 32 Monomer
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Cyan

Protein Excitation Emission Brightness* Structure Plasmids
Aquamarine 420 474 23 Monomer
ECFP 433 475 13 Monomer
Cerulean 433 475 27 Monomer
mCerulean3 433 475 32 Monomer
SCFP3A 433 474 17 Monomer
mTurquoise2 434 474 28 Monomer
CyPet 435 477 18 Monomer
AmCyan1 453 486 11 Tetramer
mTFP1 462 492 54 Monomer
MiCy (Midoriishi-Cyan) 472 495 25 Dimer
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Green

Protein Excitation Emission Brightness* Structure Plasmids
iLOV 447 497 - Monomer
AcGFP1 475 505 28 Monomer
sfGFP 485 507 54 Monomer
mEmerald 487 509 39 Monomer
EGFP or mEGFP 488 507 34 Monomer
mAzamiGreen 492 505 41 Monomer
cfSGFP2 493 517 34 Monomer
  • cfSGFP2-N - Mammalian Expression (cysteine-free SGFP2)
ZsGreen 493 505 39 Tetramer
  • pHIV-Zsgreen - Mammalian Expression (this is a bicistronic vector rather than a fusion protein vector)
  • ZsGreen1-N1 - Mammalian Expression
mWasabi 493 509 56 Monomer
SGFP2 495 512 30 Monomer
Clover 505 515 84 Monomer
mClover2 505 515 - Monomer
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Yellow

Protein Excitation Emission Brightness* Structure Plasmids
EYFP 513 527 51 Monomer
Topaz or mTopaz 514 527 57 (for Topaz) Monomer
mVenus 515 528 53 Monomer
SYFP2 515 527 69 Monomer
mCitrine 516 529 59 Monomer
YPet 517 530 80 Monomer
ZsYellow1 529 539 8 Tetramer
mPapaya1 530 541 36 Monomer
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Orange

Protein Excitation Emission Brightness* Structure Plasmids
mKusabira-Orange (mKO) 548 559 31 Monomer
mOrange 548 562 49 Monomer
mOrange2 549 565 35 Monomer
mKO2 551 565 40 Monomer
TurboRFP 553 574 62 Dimer
tdTomato 554 581 95 Tandem-dimer
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Red

Protein Excitation Emission Brightness* Structure Plasmids
mRuby2 559 600 43 Monomer
eqFP611 559 611 35 Tetramer
DsRed2 563 582 24 Tetramer
mApple 568 592 37 Monomer
mStrawberry 574 596 26 Monomer
FusionRed 580 608 18 Monomer
mRFP1 584 607 13 Monomer
mCherry 587 610 16 Monomer
mCherry2 587 610 - Monomer
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Far-Red

Protein Excitation Emission Brightness* Structure Plasmids
mCrimson3 588 615 - Monomer
HcRed1 588 618 0.3 Dimer
dKatushka 588 635 22 Dimer
mKate1.3 588 635 25 Monomer
mPlum 590 649 4 Monomer
mRaspberry 598 625 13 Monomer
TagRFP675 598 675 4 Monomer
mNeptune 600 650 13 Monomer
mNeptune2 600 650 21 Monomer
mCardinal 604 659 17 Monomer
mMaroon 610 652 - Monomer
TagRFP657 611 657 0.34 Monomer
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Near-Infrared

Protein Excitation Emission Brightness* Structure Plasmids
iRFP670 643 670 13 Dimer
iRFP682 663 682 10 Dimer
iRFP702 673 702 8 Dimer
IFP2.0 684 708 7 Monomer
iRFP (aka iRFP713) 690 713 6 Dimer
iSplit 690 713 5 Dimer
iRFP720 702 720 6 Dimer
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Long Stokes Shift

Protein Excitation Emission Brightness* Structure Plasmids
T-sapphire or mT-sapphire 399 511 26 (For T-sapphire) Monomer
mAmetrine 406 526 26 Monomer
LSSmOrange 437 572 23 Monomer
mKeima Red 440 620 3 Monomer
dKeima Red 440 616 22 Dimer
LSSmKate1 463 624 3 Monomer
LSSmKate2 460 605 4 Monomer
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Photoactivatable (e.g. off to on)

Excitation and Emission wavelengths after activation.

Protein Excitation Emission Brightness* Structure Plasmids
Phamret 458 517 14 Monomer
PA-sfGFP 485 511 - Monomer
mPA-Emerald 487 509 - Monomer
PA-GFP 504 517 14 Monomer
PATagRFP 562 595 25 Monomer
PAmCherry1 564 595 8 Monomer
PAmCherry2 570 596 13 Monomer
PAmCherry3 570 596 5 Monomer
PAmKate 568 628 5 Monomer
PAiRFP1 690 717 3 Dimer
PAiRFP2 692 719 3 Dimer
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Photoconvertible (e.g. green to red)

Protein Excitation Emission Brightness* Structure Plasmids
mEos2 506 / 571 516 / 581 47 / 30 Monomer
mEos3.2 507 / 572 516 / 580 53 / 18 Monomer
mEos4a 505 / 569 516 / 581 - Monomer
mEos4b 505 / 569 516 / 581 - Monomer
dEos 505 / 569 516 / 581 55 / 20 Dimer
tdEos 505 / 569 516 / 581 55 / 20 Tandem dimer
wtEos 505 / 569 516 / 581 50 / 23 Tetramer
wtKikGR 507 / 583 517 / 593 38 / 23 Tetramer
mKikGR 507 / 583 517 / 593 34 / 18 Monomer
Kaede 508 / 572 518 / 580 87 / 20 Tetramer
PSmOrange 548 / 634 565 / 662 58 / 9 Monomer
PSmOrange2 546 / 619 561 / 651 31 / 7 Monomer
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Photoswitchable (e.g. off to on to off)

Protein Excitation Emission Brightness* Structure Plasmids
rsTagRFP 440 / 567 585 / 585 0.02 / 4.0 Monomer
rsEmerald 491 508 - -
rsGFP1 493 510 - -
Dronpa3 503 518 19 Monomer
mGeos-M 503 / 569 514 / 581 44 Monomer
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Fluorescent Timers

Protein Excitation Emission Brightness* Structure Plasmids
Fast-FT 403(blue), 583(red) 466(blue), 606(red) 14.9(blue), 6.8(red) Monomer
Medium-FT 401(blue), 579(red) 464(blue), 600(red) 18.4(blue), 5.8(red) Monomer
Slow-FT 402(blue), 583(red) 465(blue), 604(red) 11.7(blue), 4.2(red) Monomer
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* Relative Brightness is calculated as the product of extinction coefficient and quantum yield, divided by 1000.
- Denotes unknown value.

Empty Backbones for Fluorescent Protein Fusions (Organized by Expression Species)

Organism/Expression System Plasmids
Mammalian
Insect
Zebrafish, Sea urchin, Xenopus, and C. elegans
Yeast
  • Thorn Lab Vectors - Includes tagging with a wide variety of colors, some specifically optimized for yeast
  • Lindquist Lab Vectors - Gateway cloning; includes tagging with ECFP, EYFP, DsRed, Cerulean
Bacteria
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References:

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Do you have suggestions for other plasmids or categories that should be added to this list?

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